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A simple Technique for Single-Cell Cloning of Crown Gall Tumour Tissue;PetuniaProtoplast Regeneration without Exogenous Hormones

Authors
Journal
Journal of Plant Physiology
0176-1617
Publisher
Elsevier
Publication Date
Volume
133
Issue
4
Identifiers
DOI: 10.1016/s0176-1617(88)80034-8
Keywords
  • Crown Gall Tumour
  • Protoplasts
  • Single-Cell Cloning
  • Petunia Hybrida

Abstract

Summary Protoplasts from an unselected callus line of Petunia hybrida Crown gall tumour were incubated for 3 days at high density (2 × 10 5/ml) in hormone-free medium, by which time 10 % to 20 % had divided. After plating in agarose-solidified medium at low density (8 × 10 3/ml), individual 2-cell colonies were marked and their development into independent calli monitored. The procedure demonstrates, for the first time, the ability of Crown gall tumour protoplasts to regenerate in the absence of exogenous phytohormones. Studies on regenerative ability with and without added hormones indicated the presence of transformed and untransformed cells in similar proportions in the original cell line after 3 years growth in hormonefree medium.

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